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地球科学进展  2008, Vol. 23 Issue (4): 433-438    DOI: 10.11867/j.issn.1001-8166.2008.04.0433
研究简报     
湖泊沉积物中DNA提取与PCR扩增
申慧彦1,2,李世杰1*
1. 中国科学院南京地理与湖泊研究所湖泊与环境国家重点实验室, 江苏 南京 210008;2.中国科学院研究生院, 北京 100039
Extraction and PCR Amplification of DNA from Lake Sediments
Shen Huiyan1,2,Li Shijie1
1.State Key Laboratory of Lake Science and Environment Research, Nanjing Institute of Geography and Limnology, CAS, Nanjing 210008,China; 2.Graduate School of the Chinese Academy of Sciences, Beijing 100039,China  
 全文: PDF(1128 KB)  
摘要:

对湖泊沉积物中保存的生物大分子—DNA进行研究分析,以探讨湖泊生态系统随环境改变而演变的过程。根据前人对土壤和海洋沉积物DNA的提取方法,针对湖泊沉积物的特点进行改进,在此基础上对太湖梅梁湾湖泊沉积岩芯进行DNA提取和扩增。结果表明:不同深度湖泊沉积物均能获得DNA,且纯度较高,OD260/ OD280 均大于1.4、OD260/ OD230大于1.1,可直接用于PCR(polymerase chain reaction )扩增。对微囊藻的16S rRNA基因进行PCR扩增,结果发现在前5个沉积物样品中,均得到了200 bp左右的微囊藻16S rRNA基因片段,表明微囊藻或已降解的微囊藻基因片段存在于这5个层位的湖泊沉积物中。这为利用湖泊沉积物中的生物分子来推断历史湖泊生物群落结构,判断湖泊生态演变的历史提供了新的研究思路。

关键词: 湖泊沉积物微囊藻DNA提取PCR扩增    
Abstract:

Biomacromolecules in lake sediments are very important in the indication of environmental information. DNA(Deoxyribonucleic acid) is an important biomacromolecule, and its genetic information which corresponds to organism to indicate information of biology and environmental change of lake sediments con be studied. In recent years, several approaches based on the extraction of DNA from environmental samples have been developed for the microbial diversity in soil and sea sediments. 
On the basis of previous work, an efficient method to extract DNA from lake sediments is reported. Samples from location in Lake Taihu were collected, and their genomic DNA with the size of 23kb was obtained from all sediment samples of Meiliang bay. Purification and efficiency of extracted DNA were evaluated through ultraviolet spectrum OD260/ OD280 and OD260/ OD230 ratios. Experimental results show that all samples mentioned above obtain quite purified genomic DNA with OD260/ OD280 ratios from 1.41 to 1.72, and OD260/ OD280 ratios from 1.13 to1.54. The changed regulation of DNA in different sediment depth is elucidated, the distribution of organic carbon and DNA is consistent in 15cm surface sediments, and DNA keeps low contents below 15cm. The extracted DNA is sufficient enough to be served as a template for PCR amplification of specific genetic sequences. In particular, Microcystis 16S rRNA gene fragments (200bp gene fragments) were obtained from 5 samples. 

Key words: Lake sediments    Microcystis    DNA extraction    PCR amplification.
收稿日期: 2007-12-15 出版日期: 2008-04-10
:  P512.2  
基金资助:

国家科技部重大基础研究前期研究专项“长江三角洲地区湖泊环境演变的人与自然相互作用过程研究”(编号:2004CCA02900);中国科学院知识创新工程重大项目“人类活动驱动下的长江中下游湖泊环境变化研究”(编号:KZCX1-SW-12-1)联合资助.

通讯作者: 李世杰(1955-), 男,河南偃师人,研究员, 博士生导师, 主要从事湖泊沉积与全球变化研究.     E-mail: shjli@niglas.ac.cn
作者简介: 申慧彦(1978-), 女, 山东金乡人, 博士研究生, 主要从事湖泊资源与环境研究.E-mail:huiyanshen@163.com
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引用本文:

申慧彦,李世杰. 湖泊沉积物中DNA提取与PCR扩增[J]. 地球科学进展, 2008, 23(4): 433-438.

Shen Huiyan,Li Shijie. Extraction and PCR Amplification of DNA from Lake Sediments. Advances in Earth Science, 2008, 23(4): 433-438.

链接本文:

http://www.adearth.ac.cn/CN/10.11867/j.issn.1001-8166.2008.04.0433        http://www.adearth.ac.cn/CN/Y2008/V23/I4/433

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